PicoPLEX™ WGA Kit

Whole Genome Amplification for Library Construction of Single Cells

Overview

The ability to amplify DNA to yield a highly reproducible library from a single cell is now possible! The PicoPLEX™ WGA kit with patented technology* is designed and optimized for amplification of single copy genomic DNA starting with a single cell. The easy-to-use single tube protocol reduces handling errors, dramatically improves time to results and reduces background. PicoPLEX WGA Kit can be also be used with isolated gDNA amounts ranging from less than 6 pg to 50 pg. Accepted for use in array CGH analysis as well as PCR.

PicoPLEX™ WGA Kits are used and trusted by leading providers in the IVF community such as BlueGnome for pre-implantation genetic screening and diagnostics (PGS/PGD).

PicoPLEX WGA Kit Advantages

Amplification of Single Cells is Reproducible

Flow-sorted cancer cells were amplified with the PicoPLEX WGA Kit. Each of the cells (blue lines) amplified at the same rate, and resulted in a similar, predictable yield. The sample containing 0 cells (green lines; no template control) shows very low background.

Locus-specific q-PCR was used to quantify 48 loci in independent
single-cell libraries. Data shown compares results of DNA from two
individual samples amplified with (left to right) PicoPLEX WGA Kit, GenomePLEX® and GenomiPhi™ using 10 pg of DNA. More than 90% of the product from PicoPLEX produce identifiable human sequences
and this was highly reproduced,

2. Is there a non-stick wash buffer that can be used for single cell wash prior to using PicoPLEX WGA Kit?

This has not been validated at Rubicon Genomics, but some customers have successfully used PVA (Polyvinyl Alcohol; e.g. 0.1%) in PBS to wash/collect their cells. Additionally, your washing/collecting buffer (PBS) must be also Ca- and Mg-free.

3. Are there special requirements for flow sorting?

Yes, we strongly recommend not fixing, and using light scattering or phase contrast to sort or collect samples. Microscopic/visual confirmation of successfully sorted cells can be used to optimize sorting conditions.

4. What is the sample input volume for PicoPLEX WGA Kit?

This kit will accommodate a 5 μl sample input volume per reaction (with a maximum of 2.5 μl of cell & buffer carryover).

5. Can the product of PicoPLEX WGA Kit be taken straight to NGS?

No.

6. Does Rubicon Genomics have an NGS-ready kit for single cell applications?

PicoPLEX WGA Kit Advantages

Amplification of Single Cells is Reproducible

Flow-sorted cancer cells were amplified with the PicoPLEX WGA Kit. Each of the cells (blue lines) amplified at the same rate, and resulted in a similar, predictable yield. The sample containing 0 cells (green lines; no template control) shows very low background.

Locus-specific q-PCR was used to quantify 48 loci in independent
single-cell libraries. Data shown compares results of DNA from two
individual samples amplified with (left to right) PicoPLEX WGA Kit, GenomePLEX® and GenomiPhi™ using 10 pg of DNA. More than 90% of the product from PicoPLEX produce identifiable human sequences
and this was highly reproduced,

2. Is there a non-stick wash buffer that can be used for single cell wash prior to using PicoPLEX WGA Kit?

This has not been validated at Rubicon Genomics, but some customers have successfully used PVA (Polyvinyl Alcohol; e.g. 0.1%) in PBS to wash/collect their cells. Additionally, your washing/collecting buffer (PBS) must be also Ca- and Mg-free.

3. Are there special requirements for flow sorting?

Yes, we strongly recommend not fixing, and using light scattering or phase contrast to sort or collect samples. Microscopic/visual confirmation of successfully sorted cells can be used to optimize sorting conditions.

4. What is the sample input volume for PicoPLEX WGA Kit?

This kit will accommodate a 5 μl sample input volume per reaction (with a maximum of 2.5 μl of cell & buffer carryover).

5. Can the product of PicoPLEX WGA Kit be taken straight to NGS?

No.

6. Does Rubicon Genomics have an NGS-ready kit for single cell applications?